en

B12
07P67
B12 Reagent Kit G71350R01
B7P670
Created December 2017.

07P6722
07P6732
Instructions must be carefully followed. Reliability of assay results Sample, Pre-Treatment Reagent 1, Pre-Treatment Reagent 2, and
cannot be guaranteed if there are any deviations from these Pre-Treatment Reagent 3 are combined. An aliquot of the pretreated
instructions. sample, intrinsic factor coated paramagnetic microparticles, and
ll
NAME assay diluent are combined and incubated. The B12 present in
the sample binds to the intrinsic factor coated microparticles. The
Alinity i B12 Reagent Kit mixture is washed. B12 acridinium-labeled conjugate is added to
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INTENDED USE create a reaction mixture and incubated. Following a wash cycle,
The Alinity i B12 assay is a chemiluminescent microparticle Intrinsic Pre-Trigger and Trigger Solutions are added.
Factor assay used for the quantitative determination of vitamin B12 in The resulting chemiluminescent reaction is measured as relative light
human serum and plasma on the Alinity i analyzer. units (RLUs). There is an inverse relationship between the amount of
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SUMMARY AND EXPLANATION OF THE TEST
B12 in the sample and the RLUs detected by the system optics.
For additional information on system and assay technology, refer to
Vitamin B12 (B12), a member of the corrin family, is a cofactor for the Alinity ci-series Operations Manual, Section 3.
the conversion of methylmalonyl Coenzyme-A (CoA) to succinoyl
CoA. In addition, B12 is a cofactor in the synthesis of methionine ll
REAGENTS
from homocysteine, is implicated in the formation of myelin, and, Kit Contents
along with folate, is required for DNA synthesis.1, 2 Alinity i B12 Reagent Kit 07P67
B12 is absorbed from food after binding to a protein called intrinsic NOTE: Some kit sizes are not available in all countries. Please
factor which is produced by the stomach. Causes of vitamin B12 contact your local distributor.
deficiency can be divided into three classes: nutritional deficiency,
NOTE: This product is composed of 6 components, which are
malabsorption syndromes, and other gastrointestinal causes. B12
packaged as a 2 cartridge reagent set. Both cartridges are required
deficiency can cause megaloblastic anemia (MA), nerve damage
to perform the assay.
and degeneration of the spinal cord. Lack of B12, even mild
deficiencies, damages the myelin sheath that surrounds and protects Volumes (mL) listed in the table below indicate the volume per
nerves, which may lead to peripheral neuropathy. The nerve damage cartridge set.
caused by a lack of B12 may become permanently debilitating, if 07P6722 07P6732
the underlying condition is not treated. People with intrinsic factor Tests per cartridge set 100 600
defects who do not get treatment eventually develop a MA called Number of cartridge
pernicious anemia (PA).2 2 2
sets per kit
The relationship between B12 levels and MA is not always clear in Tests per kit 200 1200
that some patients with MA will have normal B12 levels; conversely, 5.4 mL 24.8 mL
many individuals with B12 deficiency are not afflicted with MA.
Despite these complications, however, in the presence of MA (e.g., 4.9 mL 24.3 mL
elevated mean corpuscular volume (MCV)) there is usually serum 8.1 mL 42.8 mL
B12 or folate deficiency.2, 3
48.1 mL 48.1 mL
The true prevalence of B12 deficiency in the general population is
unknown but increases with age. In one study,4 fifteen percent of 5.3 mL 24.6 mL
adults older than 65 years old had laboratory evidence of vitamin 5.3 mL 24.8 mL
B12 deficiency.
A serum B12 level below the normal expected range may indicate Intrinsic Factor (porcine) coated microparticles
that tissue B12 levels are becoming depleted. However, a B12 level in borate buffer with protein (bovine) stabilizers. Minimum
in the low normal range does not ensure that B12 levels are healthy concentration: 0.1% solids. Preservative: antimicrobial agents.
and symptomatic patients should be further evaluated with tests for B12 acridinium-labeled conjugate in MES buffer.
holotranscobalamin,5 homocysteine and methylmalonic acid.6, 7 Minimum concentration: 0.7 ng/mL. Preservative: ProClin 300.
There are a number of conditions that are associated with low serum Borate buffer with EDTA. Preservative: antimicrobial
B12 levels, including iron deficiency, normal near-term pregnancy, agents.
vegetarianism, partial gastrectomy/ileal damage, celiac disease, use
of oral contraception, parasitic competition, pancreatic deficiency, 1.0 N sodium hydroxide with 0.005%
treated epilepsy, and advancing age.2, 8-11 Disorders associated with potassium cyanide.
elevated serum B12 levels include renal failure, liver disease, and Alpha monothioglycerol and EDTA.
myeloproliferative diseases.8, 12
Cobinamide dicyanide in borate buffer
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BIOLOGICAL PRINCIPLES OF THE PROCEDURE with protein (avian) stabilizers. Preservative: sodium azide.
This assay is a two-step immunoassay for the quantitative
determination of vitamin B12 in human serum and plasma using Warnings and Precautions
chemiluminescent microparticle immunoassay (CMIA) technology. •
• For In Vitro Diagnostic Use

1
Safety Precautions P337+P313 If eye irritation persists: Get medical
CAUTION: This product requires the handling of human specimens. advice / attention.
It is recommended that all human-sourced materials be considered P302+P352 IF ON SKIN: Wash with plenty of water.
potentially infectious and handled in accordance with the OSHA P332+P313 If skin irritation occurs: Get medical
Standard on Bloodborne Pathogens. Biosafety Level 2 or other advice / attention.
appropriate biosafety practices should be used for materials that P362+P364 Take off contaminated clothing and wash
contain or are suspected of containing infectious agents.13-16 it before reuse.
The following warnings and precautions apply to: / The following warnings and precautions apply to:

DANGER Contains disodium tetraborate DANGER Contains sodium hydroxide.

decahydrate. H314 Causes severe skin burns and eye
H360 May damage fertility or the unborn child. damage.
Prevention H290 May be corrosive to metals.
P201 Obtain special instructions before use. Prevention
P280 Wear protective gloves / protective P234 Keep only in original container.
clothing / eye protection. P260 Do not breathe mist / vapors / spray.
Response P264 Wash hands thoroughly after handling.
P308+P313 IF exposed or concerned: Get medical P280 Wear protective gloves / protective
advice / attention. clothing / eye protection.
Disposal Response
P501 Dispose of contents / container in P301+P330+P331 IF SWALLOWED: Rinse mouth. Do NOT
accordance with local regulations. induce vomiting.
The following warnings and precautions apply to: P305+P351+P338 IF IN EYES: Rinse cautiously with water
for several minutes. Remove contact
lenses, if present and easy to do.
Continue rinsing.
P303+P361+P353 IF ON SKIN (or hair): Take off immediately
all contaminated clothing. Rinse skin with
DANGER Contains disodium tetraborate decahydrate water / shower.
and sodium azide.
P310 Immediately call a POISON CENTER or
H360 May damage fertility or the unborn child. doctor / physician.
EUH032 Contact with acids liberates very toxic gas. P390 Absorb spillage to prevent material
Prevention damage.
P201 Obtain special instructions before use. Disposal
P280 Wear protective gloves / protective P501 Dispose of contents / container in
clothing / eye protection. accordance with local regulations.
Response
P308+P313 IF exposed or concerned: Get medical The following warnings and precautions apply to:
advice / attention.
Disposal
P501 Dispose of contents / container in
accordance with local regulations. WARNING Contains methylisothiazolones.
The following warnings and precautions apply to: H317 May cause an allergic skin reaction.
Prevention
P261 Avoid breathing mist / vapors / spray.
P272 Contaminated work clothing should not be
allowed out of the workplace.
WARNING Contains monothioglycerol. P280 Wear protective gloves / protective
clothing / eye protection.
H319 Causes serious eye irritation.
Response
H315 Causes skin irritation.
P302+P352 IF ON SKIN: Wash with plenty of water.
Prevention
P333+P313 If skin irritation or rash occurs: Get
P264 Wash hands thoroughly after handling.
medical advice / attention.
P280 Wear protective gloves / protective
P362+P364 Take off contaminated clothing and wash
clothing / eye protection.
it before reuse.
Response
Disposal
P305+P351+P338 IF IN EYES: Rinse cautiously with water
P501 Dispose of contents / container in
for several minutes. Remove contact
accordance with local regulations.
lenses, if present and easy to do.
Continue rinsing.

2
Safety Data Sheets are available at www.abbottdiagnostics.com or ll
INSTRUMENT PROCEDURE
contact your local representative. The Alinity i B12 assay file must be installed on the Alinity i analyzer
For a detailed discussion of safety precautions during system prior to performing the assay.
operation, refer to the Alinity ci-series Operations Manual, Section 8. For detailed information on assay file installation and viewing and
Reagent Handling editing assay parameters, refer to the Alinity ci-series Operations
• Upon receipt, gently invert the unopened reagent kit by rotating Manual, Section 2.
it over and back for a full 180 degrees, 5 times with green label For information on printing assay parameters, refer to the Alinity ci-
stripe facing up and then 5 times with green label stripe facing series Operations Manual, Section 5.
down. This ensures that liquid covers all sides of the bottles For a detailed description of system procedures, refer to the Alinity
within the cartridges. During reagent shipment, microparticles ci-series Operations Manual.
can settle on the reagent septum. Alternate Result Units
–– Place a check in the square on the reagent kit to indicate to Edit assay parameter "Result Units" to select an alternate unit.
others that the inversions have been completed.
Conversion formula:
• After mixing, place reagent cartridges in an upright position for
(Concentration in Default result unit) x (Conversion factor) =
48 hours before use to allow bubbles that may have formed to
(Concentration in Alternate result unit)
dissipate.
Default Result Unit Conversion Factor Alternate Result Unit
• If a reagent cartridge is dropped, place in an upright position
for 1 hour before use to allow bubbles that may have formed to pg/mL 0.7378 pmol/L
dissipate.
• Reagents are susceptible to the formation of foam and bubbles.
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SPECIMEN COLLECTION AND PREPARATION
Bubbles may interfere with the detection of the reagent level in
FOR ANALYSIS
the cartridge and cause insufficient reagent aspiration that may Specimen Types
adversely affect results. The specimen types listed below were verified for use with this
For a detailed discussion of reagent handling precautions during assay.
system operation, refer to the Alinity ci-series Operations Manual, Other specimen types and collection tube types have not been
Section 7. verified with this assay.
Reagent Storage Specimen Type Collection Tubes
Storage Maximum Additional Storage Serum Serum
Temperature Storage Time Instructions Serum separator
Unopened 2 to 8°C Until Store in upright position. Plasma Lithium heparin plasma separator
expiration If cartridge does not Sodium heparin
date remain upright, gently Dipotassium EDTA
invert the cartridge 10
times and place in an • Performance has not been established for the use of bodily
upright position for 48 fluids other than human serum and plasma.
hours before use. • The instrument does not provide the capability to verify specimen
Onboard System 30 days types. It is the responsibility of the operator to verify that the
Temperature correct specimen types are used in the assay.
Opened 2 to 8°C Until Store in upright position. Specimen Conditions
expiration If cartridge does not • Do not use:
date remain upright during –– heat-inactivated specimens
storage, discard the –– pooled specimens
cartridge. –– grossly hemolyzed specimens
Do not reuse original –– specimens with obvious microbial contamination
reagent caps or
• For accurate results, serum and plasma specimens should be
replacement caps due to
free of fibrin, red blood cells, and other particulate matter. Serum
the risk of contamination
specimens from patients receiving anticoagulant or thrombolytic
and the potential to
therapy may contain fibrin due to incomplete clot formation.
compromise reagent
performance. • To prevent cross contamination, use of disposable pipettes or
pipette tips is recommended.
Reagents may be stored on or off the system. If removed from the Preparation for Analysis
system, store reagents with new replacement caps in an upright
• Follow the tube manufacturer’s processing instructions for
position at 2 to 8°C. For reagents stored off the system, it is
collection tubes. Gravity separation is not sufficient for specimen
recommended that they be stored in their original trays or boxes to
preparation.
ensure they remain upright.
• Specimens should be free of bubbles. Remove bubbles with an
For information on unloading reagents, refer to the Alinity ci-series
applicator stick before analysis. Use a new applicator stick for
Operations Manual, Section 5.
each specimen to prevent cross-contamination.
Indications of Reagent Deterioration To ensure consistency in results, recentrifuge specimens prior to
Deterioration of the reagents may be indicated when a calibration testing if
error occurs or a control value is out of the specified range. • they contain fibrin, red blood cells, or other particulate matter.
Associated test results are invalid, and samples must be retested. NOTE: If fibrin, red blood cells, or other particulate matter are
Assay recalibration may be necessary. observed, mix by low speed vortex or by inverting 10 times prior to
For troubleshooting information, refer to the Alinity ci-series recentrifugation.
Operations Manual, Section 10.

3
Prepare frozen specimens as follows: Specimen Shipping
• Frozen specimens must be completely thawed before mixing. Package and label specimens in compliance with applicable state,
• Mix thawed specimens thoroughly by low speed vortex or by federal, and international regulations covering the transport of clinical
inverting 10 times. specimens and infectious substances.
• Visually inspect the specimens. If layering or stratification is
observed, mix until specimens are visibly homogeneous.
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PROCEDURE
• If specimens are not mixed thoroughly, inconsistent results may Materials Provided
be obtained. 07P67 Alinity i B12 Reagent Kit
• Recentrifuge specimens. Materials Required but not Provided
Recentrifugation of Specimens • Alinity i B12 assay file
• Transfer specimens to a centrifuge tube and centrifuge at a • 07P6701 Alinity i B12 Calibrators
minimum of 100 000 g-minutes. • 07P6710 Alinity i B12 Controls or other commercially available
• Examples of acceptable time and force ranges that meet this controls
criterion are listed in the table below. • 09P1540 Alinity i Multi-Assay Manual Diluent
Centrifugation time using alternate RCF values can be calculated • Alinity Pre-Trigger Solution
using the following formula: • Alinity Trigger Solution
100 000 g-minutes • Alinity i-series Concentrated Wash Buffer
Minimum Centrifugation time (minutes) =
RCF For information on materials required for operation of the instrument,
Recentrifugation Time refer to the Alinity ci-series Operations Manual, Section 1.
(Minutes) RCF (x g)* g-Minutes For information on materials required for maintenance procedures,
10 10 000 100 000 refer to the Alinity ci-series Operations Manual, Section 9.
20 5000 100 000 Assay Procedure
40 2500 100 000 For a detailed description of how to run an assay, refer to the Alinity
* To ensure consistency in results, specimens must be centrifuged ci-series Operations Manual, Section 5.
using an appropriate tube at a minimum of 2500 RCF to obtain a • If using primary or aliquot tubes, refer to the Alinity ci-series
minimum of 100 000 g-minutes. Operations Manual, Section 4 to ensure sufficient specimen is
RCF = 1.12 × rmax (rpm/1000)2 present.
RCF - The relative centrifugal force generated during • To minimize the effects of evaporation, verify adequate sample
centrifugation. cup volume is present prior to running the test.
rpm - The revolutions per minute of the rotor on which • Maximum number of replicates sampled from the same sample
the specimens are being spun (usually the digital cup: 10
readout on the centrifuge will indicate the rpm). –– Priority:
Centrifugation The time should be measured from the time the ◦◦ Sample volume for first test: 87 µL
Time - rotor reaches the required RCF or rpm to the time it ◦◦ Sample volume for each additional test from same sample
begins decelerating. cup: 37 µL
rmax - Radius of the rotor in millimeters. NOTE: If custom –– ≤ 3 hours on the reagent and sample manager:
tube adapters (i.e., adapters not defined by the ◦◦ Sample volume for first test: 150 µL
centrifuge manufacturer) are used, then the radius
◦◦ Sample volume for each additional test from same sample
(rmax) should be manually measured in millimeters
cup: 37 µL
and the RCF calculated.
–– > 3 hours on the reagent and sample manager:
g-minutes - The unit of measure for the product of RCF (× g)
and centrifugation time (minutes). ◦◦ Replace with a fresh aliquot of sample.
• Refer to the Alinity i B12 calibrator package insert and/or Alinity i
• Transfer clarified specimen to a sample cup or secondary tube B12 control package insert for preparation and usage.
for testing. For centrifuged specimens with a lipid layer, transfer • For general operating procedures, refer to the Alinity ci-series
only the clarified specimen and not the lipemic material. Operations Manual, Section 5.
Specimen Storage • For optimal performance, it is important to perform routine
Maximum maintenance as described in the Alinity ci-series Operations
Specimen Storage Manual, Section 9. Perform maintenance more frequently when
Type Temperature Time Special Instructions required by laboratory procedures.
Serum/ Room 3 days Specimens may be
Sample Dilution Procedures
Plasma temperature stored on or off the
Samples with a B12 value exceeding 2000 pg/mL (1476 pmol/L) are
clot, red blood cells, or
flagged with the code "> 2000 pg/mL" ("> 1476 pmol/L") and may
separator gel.
be diluted with either the Automated Dilution Protocol or the Manual
2 to 8°C 7 days Specimens may be Dilution Procedure.
stored on or off the Automated Dilution Protocol
clot, red blood cells, or
The system performs a 1:3 dilution of the sample and automatically
separator gel.
calculates the concentration by multiplying the result by the dilution
If testing will be delayed more than 3 days for specimens stored at factor.
room temperature or more than 7 days for specimens stored at 2 Manual Dilution Procedure
to 8°C, remove serum or plasma from the clot, red blood cells, or Suggested dilution: 1:4
separator gel and store frozen at -20°C or colder. Suggested dilution for specimens that generate repeated (2 or more)
Avoid more than 3 freeze/thaw cycles. sample aspiration errors: 1:2

4
For a 1:4 dilution, add 100 μL of the sample to 300 μL of Alinity i
Multi-Assay Manual Diluent.
For a 1:2 dilution, add 100 μL of the sample to 100 μL of Alinity i
Multi-Assay Manual Diluent.
The operator must enter the dilution factor in the Specimen or
Control tab of the Create Order screen. The system will use this
dilution factor to automatically calculate the concentration of the
sample and report the result.
The result should be > 148 pg/mL (> 109 pmol/L) before the dilution
factor is applied.
If the operator does not enter the dilution factor, the result must
be manually multiplied by the appropriate dilution factor before
reporting the result. If a diluted sample result is less than 148 pg/mL
(109 pmol/L), do not report the result. Rerun using an appropriate
dilution.
For detailed information on ordering dilutions, refer to the Alinity ci-
series Operations Manual, Section 5.
Calibration
For instructions on performing a calibration, refer to the Alinity ci-
series Operations Manual, Section 5.
Each assay control must be tested to evaluate the assay calibration.
Once a calibration is accepted and stored, all subsequent samples
may be tested without further calibration unless:
• A reagent kit with a new lot number is used.
• Daily quality control results are outside of statistically-based
quality control limits used to monitor and control system
performance, as described in the Quality Control Procedures
section of this package insert.
–– If statistically-based quality control limits are not available,
then the calibration should not exceed a 30-day limit for
recalibration frequency.
This assay may require recalibration after maintenance to critical
parts or subsystems or after service procedures have been
performed.
Quality Control Procedures
The recommended control requirement for the Alinity i B12 assay is
that a single sample of each control level be tested once every 24
hours each day of use.
Additional controls may be tested in accordance with local, state,
and/or federal regulations or accreditation requirements and your
laboratory’s quality control policy.
To establish statistically-based control limits, each laboratory should
establish its own concentration target and ranges for new control lots
at each clinically relevant control level. This can be accomplished
by assaying a minimum of 20 replicates over several (3-5) days and
using the reported results to establish the expected average (target)
and variability about this average (range) for the laboratory. Sources
of variation that should be included in this study in order to be
representative of future system performance include:
• Multiple stored calibrations
• Multiple reagent lots
• Multiple calibrator lots
• Multiple processing modules (if applicable)
• Data points collected at different times of the day
Refer to published guidelines for information or general control
recommendation, for example Clinical and Laboratory Standards
Institute (CLSI) Document C24-A3 or other published guidelines, for
general quality control recommendations.17
• If more frequent control monitoring is required, follow the
established quality control procedures for your laboratory.
• If quality control results do not meet the acceptance criteria
defined by your laboratory, sample results may be suspect.
Follow the established quality control procedures for your
laboratory. Recalibration may be necessary. For troubleshooting
information, refer to the Alinity ci-series Operations Manual,
Section 10.
5
• Review quality control results and acceptance criteria following a
change of reagent or calibrator lot.
Commercial controls should be used according to the guidelines
and recommendations of the control manufacturer. Concentration
ranges provided in the control package insert should be used only for
guidance.
For any control material in use, the laboratory should ensure that the
matrix of the control material is suitable for use in the assay per the
assay package insert.
Quality Control Guidance
Refer to “Basic QC Practices” by James O Westgard, Ph.D. for
guidance on laboratory quality control practices.18
Verification of Assay Claims
For protocols to verify package insert claims, refer to Verification of
Assay Claims in the Alinity ci-series Operations Manual.
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RESULTS
Calculation
The Alinity i B12 assay utilizes a 4 Parameter Logistic Curve fit data
reduction method (4PLC, Y-weighted) to generate a calibration and
results.
For information on alternate result units, refer to the INSTRUMENT
PROCEDURE, Alternate Result Units section of this package insert.
Flags
Some results may contain information in the Flags field. For a
description of the flags that may appear in this field, refer to the
Alinity ci-series Operations Manual, Section 5.
Measuring Interval
Measuring interval is defined as the range of values in pg/mL
(pmol/L) which meets the limits of acceptable performance for
linearity, imprecision, and bias.
The measuring interval of the Alinity i B12 assay is 148 to
2000 pg/mL (109 to 1476 pmol/L).
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LIMITATIONS OF THE PROCEDURE
• Results should be used in conjunction with other data; e.g.,
symptoms, results of other tests, and clinical impressions.
• The diagnosis of B12 deficiency cannot be solely based on
serum or plasma B12 levels. Further testing for folic acid, intrinsic
factor blocking antibodies, holotranscobalamin,5 homocysteine,
and/or methylmalonic acid is suggested for symptomatic patients
with hematological or neurological abnormalities.6, 7
• If the B12 results are inconsistent with clinical evidence,
additional testing is recommended to confirm the result.
• Hemolysis has been demonstrated to exhibit negative
interference in this B12 assay. Hemolyzed specimens should not
be analyzed.
• Specimens containing above normal protein concentrations may
generate repeated (2 or more) sample aspiration errors and
should be quantified using the Automated Dilution Protocol or
Manual Dilution Procedure (1:2).
• Heterophilic antibodies and rheumatoid factor (RF) in human
serum can react with reagent immunoglobulins, interfering with
in vitro immunoassays. Patients routinely exposed to animals or
to animal serum products can be prone to this interference and
anomalous values may be observed. Additional information may
be required for diagnosis.19
• The assay is designed to test human serum and plasma.
Specimens tested in other matrices may not give accurate
results.
• Refer to the SPECIMEN COLLECTION AND PREPARATION FOR
ANALYSIS section of this package insert for specimen limitations.
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EXPECTED VALUES Accuracy by WHO
This study was performed on the ARCHITECT i System. This study was performed on the ARCHITECT i System.
Representative performance data are provided in this section. A study was conducted to evaluate the accuracy of the ARCHITECT
Results obtained in individual laboratories may vary. B12 assay using the B12 World Health Organization International
It is recommended that each laboratory determine its own reference Standard 03/178. The assay demonstrated a -3.6% difference from
range based upon its particular locale and population characteristics. the target value of 480 pg/mL (354 pmol/L).
B12 Normals Lower Limits of Measurement
A study was performed based on guidance from Clinical and A study was performed based on guidance from CLSI EP17-A2.22
Laboratory Standards Institute (CLSI) document C28-A2.20 Serum Testing was conducted using 4 lots of the Alinity i B12 Reagent Kit
specimens from 143 individuals with normal mean corpuscular on each of 2 instruments over a minimum of 3 days. The maximum
volume, homocysteine, and folate results were assayed for B12 using observed Limit of Blank (LoB), Limit of Detection (LoD), and Limit of
the ARCHITECT B12 assay. The B12 concentration range for this Quantitation (LoQ) values are summarized below.
population was 141 to > 1218 pg/mL (104 to > 899 pmol/L) with a pg/mL pmol/L
mean of 407 pg/mL (300 pmol/L). The central 95% of the sample LoBa 83 61
population is defined below: LoDb 109 80
Expected Range 187-883 pg/mL (138-652 pmol/L) LoQc 148 109
B12 Indeterminates a The LoB represents the 95th percentile from n ≥ 60 replicates of
Levels above 300 or 400 pg/mL (221 or 295 pmol/L) are rarely zero-analyte samples.
associated with B12 deficiency induced hematological or b The LoD represents the lowest concentration at which the analyte
neurological disease, respectively. Further testing is suggested for can be detected with 95% probability based on n ≥ 60 replicates of
symptomatic patients with B12 levels between 100 and 300 pg/mL low-analyte level samples.
(74 and 221 pmol/L) (hematological abnormalities), and between 100 c The LoQ was determined from n ≥ 60 replicates of low-analyte
and 400 pg/mL (74 and 295 pmol/L) (neurological abnormalities).6, 7 level samples and is defined as the lowest concentration at which a
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SPECIFIC PERFORMANCE CHARACTERISTICS maximum allowable precision of 10 %CV and a maximum allowable
Representative performance data are provided in this section. bias of 10% were met.
Results obtained in individual laboratories may vary. Linearity
The Alinity i analyzer and the ARCHITECT i System utilize the same A study was performed based on guidance from CLSI EP06-A.23
reagents and sample/reagent ratios. This assay is linear across the measuring interval of 148 to
Unless otherwise specified, all studies were performed on the Alinity 2000 pg/mL (109 to 1476 pmol/L).
i analyzer. Analytical Specificity
Precision This study was performed on the ARCHITECT i System.
Within-Laboratory Precision The specificity of the ARCHITECT B12 assay was determined
A study was performed based on guidance from CLSI EP05-A2.21 by studying the cross reactivity with cobinamide. A human
Testing was conducted using 1 lot of the Alinity i B12 Reagent Kit, serum specimen at approximately 230 pg/mL (168 pmol/L) was
1 lot of the Alinity i B12 Calibrators, and 1 lot of the Alinity i B12 supplemented with cobinamide at 9000 pg/mL and the resulting
Controls and 1 instrument. Three controls and 2 human serum interference was 4 pg/mL (3 pmol/L).
panels were assayed in a minimum of 2 replicates at 2 separate Interference
times per day on 20 different days.
This study was performed on the ARCHITECT i System.
Within-Run Within-Laboratory
(Repeatability) (Total)a Potentially Interfering Endogenous Substances
Mean
Sample n (pg/mL) SD %CV SD %CV At the concentrations listed below, bilirubin (conjugated and
Low Control 119 251 10.4 4.2 13.5 5.4 unconjugated), total protein, and triglycerides showed less than
10% interference in the ARCHITECT B12 assay for low samples
Medium Control 120 451 17.9 4.0 21.9 4.9
(concentration range: 150 pg/mL to 250 pg/mL (111 pmol/L to
High Control 120 929 28.7 3.1 30.6 3.3
184 pmol/L)) and higher samples (concentration range: > 500 pg/mL
Serum Panel 1 118 187 12.9 6.9 14.7 7.9
(> 369 pmol/L)):
Serum Panel 2 120 1902 48.1 2.5 70.5 3.7
Potentially Interfering Substance Concentration
a Includes within-run, between-run, and between-day variability. Bilirubin < 25.1 mg/dL
Within-Run Within-Laboratory Total Protein < 12 g/dL
Mean (Repeatability) (Total)a Triglycerides < 3325 mg/dL
Sample n (pmol/L) SD %CV SD %CV
Low Control 119 185 7.7 4.2 10.0 5.4 Hemolyzed specimens should not be analyzed; refer to the
LIMITATIONS OF THE PROCEDURE section of this package insert.
Medium Control 120 333 13.2 4.0 16.2 4.9
High Control 120 685 21.1 3.1 22.6 3.3 Method Comparison
Serum Panel 1 118 138 9.5 6.9 10.8 7.9 A study was performed based on guidance from CLSI EP09-A3 using
Serum Panel 2 120 1404 35.5 2.5 52.0 3.7 the Passing-Bablok regression method.24
a Correlation Concentration
Includes within-run, between-run, and between-day variability. Units n Coefficient Intercept Slope Range
Alinity i B12 vs Serum pg/mL 126 0.99 -12.84 1.04 171-1868
ARCHITECT B12 Serum pmol/L 126 0.99 -8.92 1.04 126-1378

6
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BIBLIOGRAPHY ll
Key to Symbols
1. Lee DS, Griffiths BW. Human serum vitamin B12 assay methods - a Consult instructions for use
review. Clin Biochem 1985;18:261-266.
2. Chanarin I. Megaloblastic anaemia, cobalamin, and folate. J Clin
Pathol 1987;40:978-984. Manufacturer
3. Beuerlein FJ. Testing strategies for anemias. Lab Mgmnt 1988;23-29.
4. Pennypacker LC, Allen RH, Kelly JP, et al. High prevalence of Sufficient for
cobalamin deficiency in elderly outpatients. J Am Geriatr Soc
1992;40:1197-1204
5. Obeid R, Herrmann W. Holotranscobalamin in laboratory diagnosis Temperature limitation
of cobalamin deficiency compared to total cobalamin and
methylmalonic acid. Clin Chem Lab Med 2007;45(12):1746-1750.
6. Klee GG. Cobalamin and folate evaluation: measurement of Use by/Expiration date
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